HIF-1α介導(dǎo)鐵過載對骨髓紅系造血功能損傷的研究
[Abstract]:Aim to study the expression of hypoxia inducible factor (hypoxia inducible factor-1 偽 (HIF-1 偽) in bone marrow cells of irradiated mice with iron overload and its effect on erythroid hematopoiesis and its mechanism. Method 1. Twenty C57BL/6 mice were randomly divided into 4 groups. All the mice in the latter 3 groups were exposed to 4Gy 137Cs 緯-ray with the dose rate of 1.0 Gy / min. The mice in IO group and RAPA group were given intraperitoneal injection of iron dextran 25mg/ml (0.2ml/ only), with the exception of the latter three groups receiving 4Gy 137Cs 緯-ray irradiation at a dose of 1.0 Gy 路min ~ (- 1). The rats in Ctrl group and IR group were given intraperitoneal injection of saline (0.2ml/) for 4 weeks. The cumulative dose of iron in IR group and RAPA group was added to 50mg.RAPA group, and the body weight of rapamycin was given once a week for 4 weeks. Two months later, the cervical vertebrae were dislocated and the samples were taken. 2. Flow cytometry for detection of variable iron pool (labile iron pool,LIP in bone marrow mononuclear cells and Prussian blue staining for bone marrow cells flick to verify iron overload model. 3. Hemopoietic colony forming ability of hemopoietic stem progenitor cells was detected by semi-solid methylcellulose method: CFU-E (clony-forming units-erythroid, erythroid colony forming unit), BFU-E (burst clony-forming units-erythroid, burst erythroid hematopoietic colony formation); Flow cytometry was used to detect the number and proportion of early erythrocytes (Ter119 CD71) and late erythrocytes (Ter119 CD71-) in bone marrow. Real-time fluorescence quantitative PCR was used to detect the expression of HIF-1 偽 and HIF-1 偽 signal pathway molecules (PI3K/AKT/m TOR) m RNA) in bone marrow cells and ELISA to detect the expression level of serum EPO. Results the expression of HIF-1 偽 in IR group and IO group was (1.41 鹵0.09) times and (2.12 鹵0.17) times higher than that in Ctrl group, and the expression of HIF-1 偽 in IO group was higher than that in IR group. The percentage of early erythrocytes in IO group was lower than that in Ctrl group, and there was no significant difference (P0.05). In IR group, the percentage of late erythrocytes in IO group was lower than that in Ctrl group, and that in IO group was lower than that in IR group. There was significant difference in the number of BFU-E and CFU-E between). IR group and Ctrl group. The number of BFU-E and CFU-E in IO group was significantly lower than that in IR group (P0.05). The expression of HIF-1 偽-related signal pathway molecule (PI3K/AKT/m TOR) in both). IR group and IO group was significantly higher than that in Ctrl group (P0.05). Application of m-TOR inhibitor RAPA significantly decreased the expression of HIF-1 偽 and the expression of related signal molecules, the difference was statistically significant (P0.05). BFU-E significantly increased, the difference was statistically significant (P0.05), the number of CFU-E increased, but there was no statistical difference (P0.05); early red blood cells and late erythrocytes increased slightly, there was no statistical difference (P0.05). Conclusion 1. Iron overload can damage erythrocytic hematopoiesis of bone marrow. By constructing iron overload animal model, it was found that iron overload could affect the ability of erythropoiesis colony formation (CFU-E,BFU-E) of primordial cells and the ratio of early and late primordial erythrocytes in iron overload mice. In particular, the effect on late red blood cells was significant, and the damage was aggravated by irradiation. 2. The expression of HIF-1 偽 in bone marrow mononuclear cells of iron overloaded mice was significantly increased, suggesting that HIF-1 偽 was related to the damage of erythroid hematopoiesis of bone marrow induced by iron overload. HIF-1 偽 and its related signal transduction pathways were studied. It was found that iron overload significantly increased the PI3K,AKT,m TOR expression of signal transduction molecules in HIF-1 偽 related pathways. 3. The damage of hemopoietic function of erythroid bone marrow induced by iron overload is reversible. By inhibiting the expression of HIF-1 偽 after the application of m-TOR inhibitor rapamycin, the damage effect of iron overload on erythroid hematopoiesis of bone marrow was partially restored. This suggests that the damage of erythrocytic hematopoiesis induced by iron overload may be alleviated by inhibition of HIF-1 偽.
【學(xué)位授予單位】:天津醫(yī)科大學(xué)
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2017
【分類號】:R55
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